GSM6813254: 6547_1_6489_1_GE_sample_pool_of_24_samples_with_combinations_GCGCTTATGG_S1; Homo sapiens; RNA-Seq
Cells were harvested and diluted to a final concentration of 1,000 cells/μl in 1xPBS containing 0.04% weight/volume BSA. The Chromium Controller platform of 10X Genomics was used for single cell partitioning and barcoding. Per single cell suspension, each cell's transcriptome was barcoded during reverse transcription, pooled cDNA was amplified and Single Cell 3' Gene Expression libraries and Cell Hashing libraries via Feature barcode technology were prepared, according to the manufacturer's protocol (CG000183, CG000206 and CG000317, 10X Genomics). All libraries were quantified on a 2100 Bioanalyzer Instrument following the Agilent Technologies Protocol (Agilent DNA 7500 kit, G2938-90024). Sequence library pools were composed and quantified by qPCR, according to the KAPA Library Quantification Kit Illumina® Platforms protocol (KR0405, KAPA Biosystems) NextSeq HIGH sequencing was used for paired-end sequencing of the Single Cell 3' Gene Expression library and Cell Hashing libraries, respectively, at a sequencing depth of between 20,000-60,000 reads /cell and approximately 3,500 genes/cell. NextSeq 550 paired-end sequencing was performed using 28 cycles for Read 1, 10 cycles for Read i7, and 54 cycles for Read 2, using the NextSeq 500/550 High Output Kit v2.5 (75 Cycles) (20024906, Illumina) and NextSeq 500/550 Mid Output Kit v2.5 (150 Cycles) (20024904, Illumina).